Proteins sequestered within cellular membranes perform a variety of tasks that are critical for the viability of healthy cells. As such, membrane proteins play pivotal roles in disease and represent the majority of drug targets. Detailed structural information for membrane proteins is exceedingly scarce, due in large part to the inability to produce quantities of native, functional material that is suitable for structure determination. The objective of this program project is to develop and implement biomimetic reagents that promote stability and crystallization of membrane proteins. This objective will be met by integrating investigators from Argonne National Laboratory (multiple groups from different divisions), the University of Wisconsin, and deCODE biostructures, Inc. into a highly multidisciplinary team of scientists with complementary expertise in biochemistry, synthetic chemistry, biophysics, immunology, and materials science. The innovations to be explored within this program project include: (1) development and implementation of a membrane protein crystallization strategy that circumvents the need for detergent solubilization, (2) evaluation, design, and use of new surfactants that improve upon and replace traditional detergents, (3) selection and production of affinity reagents that assist in the stabilization, purification, and/or crystallization of functional forms of membrane proteins, and (4) the design and application of synthetic amphiphilic nanomaterials (complex fluids) for the stabilization of membrane proteins and the production of ordered arrays that yield structural information. These projects will be supported by a core facility for membrane protein production (expression, purification, and functional characterization). Initially, we will use a set of well-characterized membrane proteins as a testing ground for the development and implementation of innovative reagents, methods, and approaches to the problem of producing stable, functional membrane proteins that are suitable for crystallization and structure determination experiments in nanomaterials. In later years, techniques emerging from the project will be applied to and adjusted for a broader and more challenging range of membrane protein targets. The insights gained from early successes and failures should allow an intelligent and effective approach to a more demanding, intricate, and functionally diverse set of membrane proteins and membrane protein complexes.